青藤碱对MsPGN大鼠肾脏病理及ICAM-1表达的影响

为探讨青藤碱(Sinomenine,SIN)对实验性系膜增生性肾小球肾炎(MsPGN)的病理形态学改善及肾组织中细胞间粘附分子-1(ICAM-1)表达的影响,通过实验建立改良的慢性血清病性MsPGN动物模型,光镜观察肾小球以及肾小管-间质的病理改变情况,采用免疫组织化学法检测ICAM-1的表达并分别进行半定量分析。结果显示:光镜下,模型组与正常组相比,系膜基质指数显著升高(P<0.01),肾小球毛细血管直径明显缩小(P<0.01);青藤碱组与模型组相比,上述病理改变明显减轻,系膜基质指数显著下降(P<0.01),肾小球毛细血管直径明显改善(P<0.01)。肾组织中ICAM-1免疫组化结果显示青藤碱可明显下调ICAM-1的表达,肾组织ICAM-1的相对含量、积分光密度,均显著下降(P<0.01),青藤碱组与雷公藤多苷组相比,二者无统计学意义(P>0.05)。实验表明青藤碱能够有效减轻肾脏病理损害,抑制MsPGN大鼠肾组织ICAM-1的表达,延缓疾病进展。     

In silico analysis of stomach lineage specific gene set expression pattern in gastric cancer

Stomach lineage specific gene products act as a protective barrier in the normal stomach and their expression maintains the normal physiological processes, cellular integrity and morphology of the gastric wall. However, the regulation of stomach lineage specific genes in gastric cancer (GC) is far less clear. In the present study, we sought to investigate the role and regulation of stomach lineage specific gene set (SLSGS) in GC. SLSGS was identified by comparing the mRNA expression profiles of normal stomach tissue with other organ tissue. The obtained SLSGS was found to be under expressed in gastric tumors. Functional annotation analysis revealed that the SLSGS was enriched for digestive function and gastric epithelial maintenance. Employing a single sample prediction method across GC mRNA expression profiles identified the under expression of SLSGS in proliferative type and invasive type gastric tumors compared to the metabolic type gastric tumors. Integrative pathway activation prediction analysis revealed a close association between estrogen-α signaling and SLSGS expression pattern in GC. Elevated expression of SLSGS in GC is associated with an overall increase in the survival of GC patients. In conclusion, our results highlight that estrogen mediated regulation of SLSGS in gastric tumor is a molecular predictor of metabolic type GC and prognostic factor in GC.     

Proliferative zones in the brain of the Amur sturgeon fry. Interactions with neuromeres and migration of secondary matrix zones

Neurogenesis in the forebrain region was studied in the Amur sturgeon Acipenser schrenki fry using immunocytochemical marking of the proliferative nuclear antigen. The brain zones with high proliferative activities were located at the brain ventricle surface facing the periventricular cavity. In addition to the periventricular zone of primary proliferation, several secondary proliferative zones were found in the forebrain region of the Amur sturgeon Acipenser schrenki.     

Long-term marrow cultures: human and murine systems

The intramedullary control of marrow cell production has been a difficult area to approach experimentally. The introduction by Dr. Dexter and colleagues of long-term stromal dependent culture systems for murine marrow and the adaptation of these systems to human marrow growth have allowed for in-vitro studies of stromal dependent hemopoiesis. Despite some controversy in this area, most studies appear to show that adherent murine or human stromal cells are capable of producing a relatively large number of hemopoietic growth factors including G-CSF, GM-CSF, CSF-1, IL-6 and, at least by PCR analysis, IL-3. Other work indicates that the most primitive hemopoietic cells which appear to be multifactor responsive adhere directly to these stromal cells presumably through mediation of various adherence proteins. An early acting, multilineage factor termed hemolymphopoietic growth factor-1 (HLGF-1) has been isolated from a murine stromal cell line and may be identical to the recently described ligand for the c-kit receptor. This may represent an important early survival/maintenance factor for stem cells in this system. Studies on primitive stem cells, especially the high proliferative potential colony forming cell (HPP-CFC), indicate that they are responsive to varying combinations of growth factors and that with increasing numbers of growth factors, as studied in serum-free systems, decreasing concentrations of the factors may be biologically active. These observations altogether suggest that intramedullary hemopoiesis may be regulated by the positioning of early multifactor responsive stem cells via adherent proteins in juxtaposition to synergistically acting combinations of growth factors attached to stromal cell surfaces or the extracellular matrix.(ABSTRACT TRUNCATED AT 250 WORDS)     

Proteomic analysis of the proliferative and secretory phases of the human endometrium: protein identification and differential protein expression

Proteomic analyses of the proliferative and secretory phases of the human endometrium were carried out to identify proteins and discover differentially expressed proteins using isotope-coded affinity tags, three stages of chromatographic separation and online tandem mass spectrometry (MS/MS). From an initial list of 346 proteins identified by ProICAT, manual inspection of MS/MS spectra and confirmatory searches pared the list down to 119 positively identified proteins. Only five of the proteins showed consistent differential expression. The utility of some of these proteins as indicators of true differential expression in the endometrium is open to discussion. The two proteins with unquestionable differential expressions in the secretory endometrium are: glutamate NMDA receptor subunit zeta 1 precursor and FRAT1. Some of the proteins that show no differential expression have previously been examined in gene-expression studies with similar conclusions.     

Study of immunological and virological parameters during thalidomide treatment of SIV-infected cynomolgus monkeys

The potential therapeutic utility of thalidomide (Thd), an effective inhibitor of tumor necrosis factor (TNF)-α in vitro , was investigated in cynomolgus monkeys ( Macaca fascicularis ) at 10 months after infection with simian immunodeficiency virus (SIV). Thd-treated macaques (n=8) received an oral dose (10 mg) daily for 7 days, followed by a wash-out period of 5 weeks. A 2nd cycle of treatment was performed on the same animals at higher doses (20 mg Thd/day) for 14 days. The control monkeys (n=7) received a placebo for the same period of time. In the present study, we show that Thd, in addition to inhibiting TNF-α production after in vitro mitogen stimulation of peripheral blood mononuclear cells (PBMCs), was able to restore the proliferative responses to SIV peptides in monkeys that were infected with SIV. Interestingly, we found that such effects are associated with an increased expression of CD28 cell surface receptors on CD4⁺ T-cells paralleled by a decrease on CD8⁺ T-cells. At the same time, significant reduction in either cell-associated viral load or plasma viral RNA was not observed among the SIV-infected monkeys during the two treatment cycles, when compared with the placebo group.     

Activated mesangial cells induce glomerular endothelial cells proliferation in rat anti‐Thy‐1 nephritis through VEGFA/VEGFR2 and Angpt2/Tie2 pathway

ObjectivesWe aimed to investigate the underlying mechanism of endothelial cells (ECs) proliferation in anti‐Thy‐1 nephritis.Materials and methodsWe established anti‐Thy‐1 nephritis and co‐culture system to explore the underlying mechanism of ECs proliferation in vivo and in vitro. EdU assay kit was used for measuring cell proliferation. Immunohistochemical staining and immunofluorescence staining were used to detect protein expression. ELISA was used to measure the concentration of protein in serum and medium. RT‐qPCR and Western blot were used to qualify the mRNA and protein expression. siRNA was used to knock down specific protein expression.ResultsIn anti‐Thy‐1 nephritis, ECs proliferation was associated with mesangial cells (MCs)‐derived vascular endothelial growth factor A (VEGFA) and ECs‐derived angiopoietin2 (Angpt2). In vitro co‐culture system activated MCs‐expressed VEGFA to promote vascular endothelial growth factor receptor2 (VEGFR2) activation, Angpt2 expression and ECs proliferation, but inhibit TEK tyrosine kinase (Tie2) phosphorylation. MCs‐derived VEGFA stimulated Angpt2 expression in ECs, which inhibited Tie2 phosphorylation and promoted ECs proliferation. And decline of Tie2 phosphorylation induced ECs proliferation. In anti‐Thy‐1 nephritis, promoting Tie2 phosphorylation could alleviate ECs proliferation.ConclusionsOur study showed that activated MCs promoted ECs proliferation through VEGFA/VEGFR2 and Angpt2/Tie2 pathway in experimental mesangial proliferative glomerulonephritis (MPGN) and in vitro co‐culture system. And enhancing Tie2 phosphorylation could alleviate ECs proliferation, which will provide a new idea for MPGN treatment.     

激光显微切割/质谱联用技术在肾脏疾病诊断中的应用进展

激光显微切割(Laser microdissection,LMD)质谱(Mass spectrometry,MS)联用技术(LMD/MS)已成功应用于肾活检组织甲醛固定石蜡包埋切片的蛋白质组学研究,提高了某些肾脏病的诊断水平,显示出较好的临床应用前景。文中就LMD/MS蛋白质组学技术的原理、方法及该技术在肾淀粉样变性、膜增殖性肾小球肾炎等肾脏疾病的发病机制及诊断分型的应用进展进行综述。     

肠道病毒71型在RD细胞和Vero细胞中的增殖动力学特征

目的:利用噬斑法比较肠道病毒71型(EV71)在RD细胞和Vero细胞中的增殖动力学特征。方法:首先探讨培养基类型、羟乙基哌嗪乙磺酸(HEPES)、胎牛血清(FBS)、牛血清白蛋白(BSA)及甲基纤维素(MC)含量对EV71噬斑形成的影响,得到最适营养覆盖物配比;进一步,EV71以感染复数(MOI)为0.1分别接种RD细胞和Vero细胞,收集接种后不同时间点的细胞培养液,噬斑法测定各时间点培养液上清中的病毒滴度,并绘制log2(病毒滴度)-时间图,对比分析EV71在2种细胞中的增殖动力学特征。结果:终浓度含1%MC和2%FBS的MEM(1×)或DMEM(1×)为EV71噬斑形成的最适营养覆盖物;EV71在RD细胞和Vero细胞中的增殖周期均约为12 h,MOI=0.1时,EV71在RD细胞中的增殖活动较Vero细胞中活跃,增殖效率比Vero细胞中高2个数量级。结论:用RD细胞扩增EV71比Vero细胞更具优势。